Fig. 4

Mechanism of action of AVEC: anti-HER-2 × HBsAg in ovarian cancer cells’ killing. (a) The patients’ ovarian cancer cells were labeled for 6 h at 37 °C with 0.3 mg/ml trastuzumab or anti-HER-2 × HBsAg (B) in full erythrocytes’ free blood from the HBV-vaccinated patients with the anti-HBV adjusted to 10.0 IU/ml. That was followed by labeling with anti-phosphatidylserine (anti-PS) or propidium iodide (PI) or anti-genomic DNA (antigDNA). While most of the cells demonstrate a flip of phosphatidylserine, only some of them have compromised membranes’ permeability for makers of intranuclear, genomic DNA. The assays were repeated four times. The data presented are representative for all acquired. (b) Effects of AVEC upon the ovarian cancer cells’ growth was studied by incorporation of tritium tagged thymidine. AVEC inflicted statistically significant higher impact upon the cells’ growth, when compared to isotype antibodies. AVEC had negligible effects upon HAE and HOSE cells. (c) Treatment induced apoptosis was evaluated by labeling with anti-phosphatidylserine (anti-PS) superparamagnetic antibodies and measuring relaxivity in NMR. Anti-HER-2 naked antibodies resulted in approximately 40% of cells being apoptotic. Treatment with the AVEC resulted in the number of apoptotic cells more than doubled. (d) Treatment induced necrosis was evaluated by labeling with anti-genomic DNA (anti-gDNA) superparamagnetic antibodies and measuring relaxivities in NMR. Anti-HER-2 naked antibodies resulted in approximately 10% of cells being necrotic. Treatment with the AVEC resulted in the number of necrotic cells nearly tripled. I isotype antibody. These assays were repeated four times. The data presented are representative for all performed